Preparation of aqueous clear solution dosage forms with bile acids

ABSTRACT

Compositions for pharmaceutical and other uses comprising clear aqueous solutions of bile acids which do not form any detectable precipitates over selected ranges of pH values of the aqueous solution and methods of making such solutions. The compositions of the invention comprise water; a bile acid in the form of a bile acid, bile acid salt, or a bile acid conjugated with an amine by an amide linkage; and either or both an aqueous soluble starch conversion product and an aqueous soluble non-starch polysaccharide. The composition remains in solution without forming a precipitate over a range of pH values and, according to one embodiment, remains in solution for all pH values obtainable in an aqueous system. The composition, according to some embodiments, may further contain a pharmaceutical compound in a pharmaceutically effective amount. Non-limiting examples of pharmaceutical compounds include insulin, heparin, bismuth compounds, amantadine and rimantadine.

This application claims the benefit of provisional application No.60/180,268, filed Feb. 4, 2000 and is a continuation in part ofapplication Ser. No. 09/357,549 filed Jul. 20, 1999, now U.S. Pat. No.6,251,428, which claims the benefit of provisional application No.60/094,069, filed Jul. 24, 1998, all of which are incorporated herein byreference.

BACKGROUND OF THE INVENTION

Bile acids salts which are organic acids derived from cholesterol arenatural ionic detergents that play a pivotal role in the absorption,transport, and secretion of lipids. In bile acid chemistry, the steroidnucleus of a bile acid salt has the perhydrocyclopentano phenanthrenenucleus common to all perhydrosteroids. Distinguishing characteristicsof bile acids include a saturated 19-carbon sterol nucleus, abeta-oriented hydrogen at position 5, a branched, saturated 5-carbonside chain terminating in a carboxylic acid, and an alpha-orientedhydroxyl group in the 3-position. The only substituent occurring in mostnatural bile acids is the hydroxyl group. In most mammals the hydroxylgroups are at the 3, 6, 7 or 12 positions.

The common bile acids differ primarily in the number and orientation ofhydroxyl groups on the sterol ring. The term, primary bile acid refersto these synthesized de novo by the liver. In humans, the primary bileacids include cholic acid (3α, 7α12α-trihydroxy-5β-cholanic acid) (“CA”)and chenodeoxycholic acid (3α, 7α-dihydroxy-5β-cholanic acid) (“CDCA”).Dehydroxylation of these bile acids by intestinal bacteria produces themore hydrophobic secondary bile acids, deoxycholic acid (3α,12α-dihydroxy-5β-cholanic acid) (“DCA”) and lithocholic acid(3α-hydroxy-5β-cholanic acid) (“LCA”). These four bile acids CA, CDCA,DCA, and LCA, generally constitute greater than 99 percent of the bilesalt pool in humans. Secondary bile acids that have been metabolized bythe liver are sometimes denoted as tertiary bile acids.

Keto-bile acids are produced secondarily in humans as a consequence ofoxidation of bile acid hydroxyl groups, particularly the 7-hydroxylgroup, by colonic bacteria. However, keto-bile acids are rapidly reducedby the liver to the corresponding α or β-hydroxy bile acids. Forexample, the corresponding keto bile acid of a CDCA is 7-ketolithocholic acid and one of its reduction products with thecorresponding β-hydroxy bile acid is ursodeoxycholic acid(3α-7β-dihydroxy-5β-cholanic acid) (“UDCA”), a tertiary bile acid.

UDCA, a major component of bear bile, has been used for the treatment ofand the protection against many types of liver disease for a little over70 years as a major pharmaceutical agent. Its medicinal uses include thedissolution of radiolucent gall stones, the treatment of biliarydyspepsias, primarily biliary cirrhosis, primary sclerosingchoplangitis, chronic active hepatitis and hepatitis C. In othermammalian species, bile acids containing a 6β-hydroxyl group, which arefound in rats and mice, are known as muricholic acid; 6α-hydroxy bileacids produced by swine are termed hyocholic acid and hyodeoxycholicacids. 23-hydroxy bile acids of aquatic mammals are known as phocecholicand phocedeoxycholic acids.

Typically, more than 99 percent of naturally occurring bile saltssecreted into human bile are conjugated. Conjugates are bile acids inwhich a second organic substituent (e.g. glycine, taurine, glucuronate,sulfate or, rarely, other substituents) is attached to the side chaincarboxylic acid or to one of the ring hydroxyl groups via an ester,ether, or amide linkage. Therefore, the ionization properties ofconjugated bile acids with glycine or taurine are determined by theacidity of the glycine or taurine substituent.

Free, unconjugated, bile acid monomers have pK_(a) values ofapproximately 5.0. However, pK_(a) values of glycine conjugated bileacids are on average 3.9, and the pK_(a) of taurine conjugate bile acidsare less than 1.0. The effect of conjugation, therefore, is to reducethe pK_(a) of a bile acid so that a large fraction is ionized at anygiven pH. Since the ionized salt form is more water soluble than theprotonated acid form, conjugation enhances solubility at a low pH. Freebile acid salts precipitate from aqueous solution at pH 6.5 to 7. Incontrast, precipitation of glycine conjugated bile acid occurs only atpH of less than 5. Taurine conjugated bile acids remain in aqueoussolution under very strongly acidic conditions (lower than pH 1).However, in the gastric pH range, certain bile acids such as UDCA andCDCA are no longer soluble.

Conjugation of the side chain of a bile acid with glycine or taurine haslittle influence on the hydrophobic activity of fully ionized bilesalts. More hydrophobic bile salts exhibit greater solubilizing capacityfor phospholipid and cholesterol and are consequently better detergents.More hydrophobic bile salts are also more injurious to variousmembranes, both in vivo and in vitro.

Natural bile salt pools invariably contain multiple bile acid salts.Mixtures of two or more bile salts of differing hydrophobic activity maybehave as a single bile salt of an intermediate hydrophobic activity. Asa result, detergent properties and the toxicity of mixtures of two bileacids of differing hydrophobic activity often are intermediate betweenthe individual components. Biologic functions and biologic properties ofbile acids resulting from their amphiphillic properties are as follows:

-   1. Ursodeoxycholic acid is a useful immuno-modulating agent.-   2. Ursodeoxycholic acid inhibits deoxycholic acid-induced apoptosis    by modulating mitochondrial transmembrane potential and reactive    oxygen species production.-   3. Ursodeoxycholic acid inhibits induction of nitric oxide synthase    (NOS) in human intestinal epithelial cells and in vivo.-   4. The hydrophilic nature of ursodeoxycholic acid confers    cytoprotection in necroinflammatory diseases of the liver.-   5. Ursodeoxycholic acid significantly improves transaminases and    cholestatic enzymatic indices of liver injury in chronic hepatitis.-   6. Bile acids substantially inhibit the growth of H. pylori.-   7. Ursodeoxycholic acid is the most potent pepsin inhibitor among    bile acids.-   8. High levels of bile acids remarkably inhibit the proliferation of    hepatitis C virus.-   9. Ursodeoxycholic acid has cell membrane stabilizing properties.-   10. Ursodeoxycholic acid alleviates alcoholic fatty liver.-   11. Ursodeoxycholic acid has a vasodilative effect on the systemic    vascular bed but altered neither pulmonary vascular function nor    cardiac functions.-   12. Bile acid synthesis from cholesterol is one of the two principal    pathways for the elimination of cholesterol from the body.-   13. Bile flow is generated by the flux of bile salts passing through    the liver. Bile formation represents an important pathway for    solubilization and excretion of organic compounds, such as    bilirubin, endogenous metabolites, such as emphipathic derivatives    of steroid hormones; and a variety of drugs and other xenobiotics.-   14. Secretion of bile salts into bile is coupled with the secretion    of two other biliary lipids, phosphatidylcholine (lecithin) and    cholesterol. Coupling bile salt output with the lecithin and    cholesterol output provides a major pathway for the elimination of    hepatic cholesterol.-   15. Bile salts, along with lecithin, solubilize cholesterol in bile    in the form of mixed micelles and vesicles. Bile salt deficiency,    and consequently reduced cholesterol solubility in bile, may play a    role in the pathogenesis of cholesterol gallstones.-   16. Bile acids are thought to be a factor in the regulation of    cholesterol synthesis. At present, it is not certain whether they    regulate the cholesterol synthesis by acting directly on the    hydroxymethylglutaryl-coenzyme A (HMG-CoA) reductase or indirectly    by modulating the cholesterol absorption in the intestine.-   17. Bile salts in the enterohepatic circulation are thought to    regulate the bile acid synthesis by suppressing or derepressing the    activity of cholesterol 7-hydroxylase, which is the rate-limiting    enzyme in the bile acid biosynthesis pathway.-   18. Bile acids may play a role in the regulation of hepatic    lipoprotein receptors (apo B.E.) and consequently may modulate the    rate of uptake of lipoprotein cholesterol by the liver.-   19. In the intestines, bile salts in the form of mixed micelles    participate in the intraliminal solubilization, transport, and    absorption of cholesterol, fat-soluble vitamins, and other lipids.-   20. Bile salts may be involved in the transport of calcium and iron    from the intestinal lumen to the brush border.

Recent drug delivery research concerning the characteristics andbiofunctions of naturally occurring bile acid as an adjuvant and/or acarrier has focused on the derivatives and analogs of bile acids andbile acids themselves as novel drug delivery systems for delivery to theintestinal tract and the liver. These systems exploit the activetransport mechanism to deliver drug molecules to a specific targettissue by oral or cystic administration. Thus, if bile acids or bileacid derivatives are rapidly and efficiently absorbed in the liver and,consequently, undergo enterohepatic cycling, many potential therapeuticapplications are foreseen including the following: improvement of theoral absorption of an intrinsically, biologically active, but poorlyabsorbed hydrophillic and hydrophobic drug; liver site-directed deliveryof a drug to bring about high therapeutic concentrations in the diseasedliver with the minimization of general toxic reactions elsewhere in thebody; and gallbladder-site delivery systems of cholecystographic agentsand cholesterol gallstone dissolution accelerators. As an example, in1985, Drs. Gordon & Moses et al. demonstrated that a therapeuticallyuseful amount of insulin is absorbed by the nasal mucosa of human beingswhen administered as a nasal spray with common bile salts such as DCA,UDCA, CDCA, CA, TUDCA, and TCDCA. See Moses, Alan C., et al., Diabetesvol. 32 (November 1983) 1040-1047; Gordon, G. S., et al., Proc. Nat'lAcad. Sci. USA, vol. 82 (November 1985) 7419-7423. In their experiment,bile acids produced marked elevations in serum insulin concentration,and about 50 percent decreases in blood glucose concentrations. However,this revolutionary nasal spray solution dosage form with bile acids(salts) as an adjuvant could not be developed further andcommercialized, because the nasal spray solution must be preparedimmediately prior to use due to the precipitation of bile acid salt andthe instability of insulin at pH levels between 7.4 and 7.8. Moreover,as indicated in this disclosure, ursodeoxycholic acid as an adjuvantcould not be used because of its insolubility at pH between 7.4 and 7.8.

Bile acid salts and insulin, thus, appear to be chemically andphysically incompatible. The pH of commercial insulin injectionsolutions is between 2.5 and 3.5 for acidified dosage forms and isbetween 7.00 and 7.4 for neutral dosage forms. Dosage forms of bile acidsalts prepared by conventional techniques have been unable to overcomeproblems with bile precipitation at these pH levels and insulin isunstable at a pH of 7.4 or higher. Therefore, safe and efficientpreparations of any solution dosage forms of insulin with bile acid(salt) are not commercially available at this time.

Heparin, a most potent anticoagulant, is widely used in the treatment ofand in the prevention of thromboembolism. However, heparin treatment isusually limited to hospitalized patients since this drug is given onlyby injection. Alternate routes which have been attempted are anintrapulmonary spray, suppositories, and enema. According to numerouspublications, for heparin absorption through the gastrointestinal mucosato be facilitated, the preparations should be in acidic condition.According to Dr. Ziv, Dr. Eldor et al., heparin was absorbed through therectal mucosa of rodents and primates only when administered insolutions containing sodium cholate or sodium deoxycholate. See Ziy E.et al., Biochemical PharmacoloMy, vol. 32, No. 5, pp. 773-776 (1983).However, heparin is only stable under acidic conditions. Bile acids areparticularly not soluble in acidic conditions. Therefore, due to theirincompatible characteristics, commercial dosage forms of bile acids withheparin are not presently available.

Drug delivery systems involving bile acids can provide liver-specificdrug targeting which is of major interest for drug development sincestandard pharmacological approaches to liver diseases have beenfrustrated by the inadequate delivery of active agents into liver cellsas well as non specific toxicity towards other organs. For example, theliver-specific delivery of a drug is necessary for inhibitors ofcollagen synthesis for the treatment for liver fibrosis in order toavoid unspecific and undesired side-effects in extrahepatic tissues.Furthermore, for the treatment of cancer of the biliary system, highdrug levels must be achieved in the liver and the biliary system,whereas in extrahepatic tissues low drug concentrations are desired tominimize the cytoxicity of the cytostatics to normal non-tumor cells.Dr. Kramer, Dr. Wess et al. demonstrate that hybrid molecules formed bycovalent linkages of a drug to a modified bile acid molecule arerecognized by the Na+-dependent bile acid uptake systems in the liverand the ileum. See U.S. Pat. No. No. 5,641,767. Even if bile acid saltsand their derivatives act as shuttles for specific delivery of a drug tothe liver, as already mentioned above, there are enormous risks to thedevelopment of the derivatives of bile acids or bile acid salts ascarriers because new derivatives of bile acids or bile acid salts formedby covalent linkages of a drug to bile acid must be tested for itspharmacology, toxicity and clinical effectiveness. Thus, the developmentof preparations in which a drug can be absorbed with bile acids or bileacid salts from the places which contain the excessive bile acids in theintestine is far easier and far more valuable than the development ofthe new bile acid derivatives because less testing is required.

In spite of the extremely valuable therapeutic activities and the longhistoric medical uses of bile acids as therapeutically active agents andas carriers and/or adjuvants based on the already mentioned biologicalproperties and fuictions of bile acids, the commercial administration ofbile acids is limited to pharmaceutical formulations with a solid formof bile acid which are in tablet, capsule and suspension. This is due tothe insolubility of bile acid in aqueous media at pH from approximately1 to 8. This is also due to bile's extremely bitter taste and equallybitter after-taste which lasts several hours. Ursodeoxycholic acid,chenodeoxycholic acid, and lithocholic acid are practically insoluble inwater. Deoxycholic acid and cholic acid have solubilities of 0.24 g/L,and 0.2 g/L, respectively. Tauroursodeoxycholic acid,taurochenodeoxycholic acid, and taurocholic acid are insoluble inhydrochloric acid solution. The few aqueous dosage forms that areavailable are unstable, and have very limited uses because of pH controland maintenance problems. Moreover, some commercial pharmaceuticaldosage forms of bile acids have been shown to have scant bioavailabilityas described in European Journal of Clinical Investigation (1985) 15,171-178. Bile acid, especially ursodeoxycholic acid is poorly soluble inthe gastro-duodeno j ejunal contents of fasted subjects. From 21% to 50%of the ingested doses were recovered in solid form because of theunpredictable variations in the very slow progressive solubilization ofsolid ursodeoxycholic acid in the gastrointestinal track. Bile acids,particularly ursodeoxycholic acid, deoxycholic acid, chenodeoxycholicacid, cholic acid, hyodeoxycholic acid, 7-keto lithcholic acid,tauroursodeoxycholic acid, and taurochenodeoxycholic acid among others,are especially insoluble in the gastric juices and in aqueoushydrochloric acid solution. However, the solubility of bile acidsincrease with the increase of the pH in the intestine very slowly andincompletely, and eventually the bile acids become soluble at pH between8 and 9.5.

To overcome this slow and inefficient absorption process in theintestine due to the incomplete and slow solubilization of bile acids,many newly developed pharmaceutical formulations have been prepared,such as delayed release dosage forms with water soluble solid bile acidswhich are often strongly alkaline. These newly developed pharmaceuticaldosage forms are enterosoluble-gastroresistant. Theseenterosoluble-gastroresistant dosage forms remain intact in gastricjuices in the stomach, but are dissolved and release the stronglyalkaline solid bile salts of the formulations at the targeted area,within a limited time once they reach the small intestine.

These types of dosage forms, of course, showed better bioavailabilitythan presently commercialized dosage forms as described in U.S. Pat. No.5,380,533. However, it is extremely difficult and very costly to preparethe precise delayed release dosage forms which can releasetherapeutically active components by disintegration, dissolution anddiffusion at the desired area within a limited time. According to U.S.Pat. No. 5,302,398, the absorption test of the gastroresistantenterosoluble dosage forms of bile acids, particularly ursodeoxycholicacid in man show that its absorption increases a value of about 40percent in comparison with administering the same amount in currentcommercial dosage forms. Its maximum hematic concentrations are onaverage three times higher, and are reached faster than with thecommercial formulations. Any dosage forms of bile acid formula must becapable of releasing bile acids in a known and consistent mannerfollowing administration to the patient. Both the rate and the extent ofrelease are important, and should be reproducible. Ideally, the extentof release should approach 100 percent, while the rate of release shouldreflect the desired properties of the dosage form.

It is a well-known fact that solution dosage forms of drugs showsignificantly improved rates and extents of absorption, compared to thesame drug formulated as a tablet, capsule, or suspension. This isbecause solution dosage forms are chemically and physically homogeneoussolutions of two or more substances. Moreover, the specially designedsolution dosage forms which can maintain the solution systems withoutbreaking down under any pH conditions are ready to be diffused in thedesired area for immediate and complete absorption, whereas tablets,capsules or delayed release formulations must invariably undergodisintegration, dissolution and diffusion at the desired area within alimited time. Unpredictable variations in the extent and rate of releaseof bile acids by the disintegration, dissolution and diffusion ofdelayed or immediate release dosage forms having pH-dependentinstability result in the slow and inefficient bile absorption andreduced bioavailability.

The luminal surface of the stomach is coated with a thick layer ofprotective mucus. The mucus gel coating maintains a pH gradient from theintraluminal compartment to the apical membrane and is believed tocontribute to the phenomenon of cytoprotection. H. pylori infectionoccurs on the luminal surface of the stomach mucosa within the mucus, onthe epithelial surface, and within the gastric pints. Bacterial enzymesare believed to degrade the mucus glycoprotein network and reduce thepolymers to monomers (or subunits) such that the mucus can no longerexist as a gel. In addition, mucinogenesis is reduced and the mucosabecomes susceptible to the erosive effects of acid. This condition maylead to gastritis and peptic ulcers.

Bismuth compounds have gained increasing interest in the therapeutictreatment of gastro-duodenal disorders and especially in the eradicationof Helicobacter pylori, a bacterium thought to be involved in theetiology of the disease. Many oral preparations of bismuth have beenused. The various preparations appear to differ in clinical efficacy aswell as pharmacokinetics. The inorganic salts used have includedsubnitrate, subcarbonate, subgallate, tartarate, citrate andsubsalicylate. The commercial preparations have generally been availableover the counter and have often contained other compounds in addition tothe bismuth salt. The commercial preparations have been usedsuccessfully in the treatment of both gastric and duodenal ulcerdisease. These preparations have proved as effective as the histamine H2antagonists in the treatment of gastric and duodenal ulcers and havebeen associated with lower relapse rates after cessation of therapy. Thelower relapse rate after initial healing with bismuth preparations havebeen attributed to its ability to eradicate H. pylori and to moderatethe gastroduodenitis associated with infection by this organism.Long-term eradication of H. pylori is more likely when bismuthpreparation is administered along with antibiotics or antiseptics (localdelivery) such as bile acids.

A variety of antibiotics and antiseptics display good activity againstH. pylori in vitro. Yet when tested as single agents in clinicalstudies, they do not succeed in eradicating the organism. Failure oftherapy and relapse are very common. The reason for this discrepancybetween in vitro and clinical results has not been established. Possibleexplanations are poor penetration of the compounds into gastric mucus,destruction at acid pH, insolubility in acidic environment, andcombinations thereof. Consequently, administration of high doses ofantimicrobial agents on a daily basis is necessary for H. pylorieradication. The efficacy of this course of therapy is hindered by poorpatient compliance due to adverse effects such as diarrhea, nausea,retching and breakdown of normal intestinal flora.

Another reason for incomplete eradication may be that the residence timeof antimicrobial agents in the stomach is so short that effectiveantimicrobial concentrations cannot be achieved in the gastric mucouslayer or epithelial cell surfaces where H. pylori exist. Therefore,eradication of H. pylori may be better achieved by a therapy thatimproves antimicrobial agent delivery for topical activity andabsorption for systemic activity. However, no in vivo eradication trialswith dosage forms that prolong the gastric residence times for topicalactivity and have the high absorption rate in the gastro-intestinaltract have been reported. The best results so far have been achievedwith the combination of a non-absorbed agent with topical activity,colloidal bismuth compounds, and a well-absorbed agent with systemicactivity, amoxicillin (Van Caekemberghe and Breyssens, 1987,Antimicrobial Agent and Chemotherapy, pp 1429-1430). But clearly,therapy for H. pylori infections is still suboptimal.

In addition to a bactericidal effect, some bismuth compounds haveprofound effects on some of the pathogenic mechanisms whereby H. pyloridamage the mucosa. Bismuth compounds are potent and non-specific enzymeinhibitors. In vitro studies have suggested that these compounds mayinhibit bacterial enzymes, including lipases, proteases, andglycosidases synthesized by H. pylori. Inhibition of bacteria enzymesand maintenance of an intact viscoelastic gel coating is thought to berelated to the therapeutic action of bismuth compounds for H. pyloriassociated gastritis and peptic ulcers.

Bismuth compounds block the adhesion of H. pylori to epithelial cells.Shortly after oral administration of these compounds, the organisms werelocated inside, rather than underneath, the mucus gel. This was thoughtto result from loss of adherence to the apical membrane of the gastricepithelial cells. Bismuth, in the form of electron dense bodies, wasseen to be deposited on the surface and within the bacterial cell. Butunfortunately, intramucus bismuth concentrations often fall below themean inhibitory concentration of bismuth compounds for H. pylori,because of the diluting effects of food, long disintegration time ofcommercial tablets at pH<1.1, and bismuth precipitation due toinsolubility in acidic environment. Additionally, H. pylori inactivatedby exposure to growth-inhibiting concentrations of bismuth compounds canremain viable for several hours and, therefore is capable of resumingnormal growth when bismuth is removed.

The inhibitory factor(s) in bile was markedly reduced by acidificationfollowed by centrifugation to remove precipitated glycine-conjugatedbile acids and was completely eliminated by use of the bileacid-sequestering agent cholestyramine. These results are consistentwith the notion that acidic conditions in the duodenal bulb would serveto precipitate inhibitory bile acids (or other inhibitory substances)and allow H. pylori to grow in an otherwise hostile environment. Theobservations related to reflux gastritis by D. Y. Graham (Osato et al.,1999, Digestive Diseases and Sciences 44(3):462-464) can be extended topossibly understand the relation between acid secretion and duodenalulcer. Low pH in the duodenal bulb in patients with duodenal ulcerdisease associated with high acid secretion, rapid gastric emptying, andlocal production of acid would both promote development of gastricmetaplasia and precipitate the deleterious glycine-conjugated bileacids, allowing H. pylori to colonize and thrive.

SUMMARY OF THE INVENTION

The present invention relates to a composition which comprises (1) abile acid, its derivative, its salt, or its conjugate with an amine, (2)water, and (3) a sufficient quantity of an aqueous soluble starchconversion product such that the bile acid and the starch conversionproduct remain in solution at any pH within a selected pH range.

The invention further relates to a composition which comprises (1) abile acid, its derivative, its salt, or its conjugate with an amine, (2)water, and (3) a sufficient quantity of an aqueous soluble non-starchpolysaccharide such that the bile acid and the polysaccharide remain insolution at any pH within a selected pH range.

The invention further relates to a pharmaceutical composition whichcomprises (1) a bile acid, its salt, or its conjugate with an amine, (2)water, (3) a pharmaceutical compound in a pharmaceutically appropriateamount, and (4) a sufficient quantity of an aqueous soluble starchconversion product or an aqueous soluble non-starch polysaccharide suchthat the bile acid, the pharmaceutical compound, and the carbohydrateremain in solution at any pH level within a selected pH range.

The invention further relates to solution dosage forms of bile acidcompositions. Advantages of these solution dosage forms include improvedbioavailability and absorbability of a bile acid. Additional advantagesof solution dosage forms include improved bioavailability andabsorbability of a pharmaceutical compound.

In some embodiments of the invention, a composition is provided whichcomprises (1) a bile acid, its derivative, its salt, or its conjugatewith an amine, (2) water, and (3) a sufficient quantity of carbohydratesuch that the bile acid component and the carbohydrate remain insolution at any pH within a selected pH range, wherein the carbohydrateis a combination of an aqueous soluble starch conversion product and anaqueous soluble non-starch polysaccharide. In embodiments containingboth soluble non-starch polysaccharide and high molecular weight starchconversion product, the amounts of each are such that when combinedtogether in the composition they are sufficient to allow the bile acidcomponent, the high molecular weight starch conversion product, thesoluble non-starch polysaccharide and the pharmaceutical compound, ifany, to remain in solution at any pH within a selected pH range.

In some embodiments of the invention, a combination therapy compositionis provided which may increase intensity of response to or efficacy of apharmaceutical. Such a composition may permit administration of lowerdosages of a pharmaceutical compound, attack a disease complex atdifferent points, affect elimination and/or alter absorption of apharmaceutical compound. Such a composition may lead to or contribute toa reduction in toxicity and/or side effects of a pharmaceutical.

The invention further relates to a composition which comprises (1) abile acid, its salt, or its conjugate with an amine, (2) water, (3) anaqueous soluble bismuth compound, and (4) a sufficient quantity of anaqueous soluble starch conversion product or an aqueous solublenon-starch polysaccharide such that the bile acid, such that the bileacid, bismuth, and carbohydrate remain in solution at any pH levelwithin a selected pH range.

The invention further relates to a method of treating or preventing ahuman or animal disease comprising administration of a composition ofthe invention.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1: Graph of blood serum—concentration of UDCA (squares) and GUDCA(triangles) versus time following administration of dosage formulationsaccording to Examples II and VI and Table 4.

FIG. 2: Graph of blood serum concentration of UDCA versus time followingadministration of dosage formulations of the bile acid according toExamples III and VI and Table 4.

FIG. 3: Diagram of the mean(n=5) for group I for pharmacokineticparameters of UDCA in human after an oral administration of liquidformulation of UDCA prepared according to Example IX without bismuth.

FIG. 4: Diagram of the mean(n=5) for group II for pharmacokineticparameters of UDCA in human after an oral administration of liquidformulation of UDCA prepared according to Example IX.

FIG. 5A. Transmission electron micrograph of H. pylori cultured fromColumbia medium.

FIG. 5B. Transmission electron micrograph of H. pylori 48 hrs afterbeing treated with UDCA & bismuth citrate prepared according to ExampleIX.

FIG. 5C. Transmission electron micrograph of H. pylori 72 hrs afterbeing treated with UDCA & bismuth citrate.

FIG. 6: NMR data for UDCA in a liquid formulation dosage form preparedaccording to Example III without preservatives, flavoring agent, andsweetener.

FIG. 7: HPLC trace of UDCA in a liquid formulation dosage form preparedaccording to Example III without preservatives, flavoring agent, andsweetener.

FIG. 8: HPLC trace of a UDCA standard.

FIG. 9: H. pylori culture method.

FIG. 10: H. pylori culture method.

FIG. 11: H. pylori culture method.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to an aqueous solution comprising (i) oneor more soluble bile acids, aqueous soluble bile acid derivatives, bileacid salts, or bile acid conjugated with an amnine, (collectively “bileacid”), (ii) water, and (iii) one or more aqueous soluble starchconversion products or aqueous soluble non-starch polysaceharide in anamount sufficient to produce a solution which does not form aprecipitate at any pH within a desired pH range. In a preferredembodiment of the invention, the bile acid and the carbohydrate do notprecipitate between about pH 1 and about pH 10, more preferably betweenabout pH 1 and about pH 14, and most preferably at all pH valuesobtainable in an aqueous system. In some embodiments of the invention, abile acid remains dissolved under acidic conditions as a free bile acidin spite of the general insolubility of bile acids under acidicconditions. In some embodiments of the invention, the composition may beused as a pharmaceutical formulation wherein the pharmaceutical compoundremains in solution without precipitation at prevailing pH levels in themouth, stomach and intestines. The composition may contain a bile acidor its salt which itself has pharmaceutical effectiveness. Formulationsof the invention may act as a carrier, an adjuvant or enhancer for thedelivery of a pharmaceutical material which remains dissolved in thecomposition of the invention across the desired pH range. In someembodiments of the invention, a non-bile acid pharmaceutical is usedthough not in solution.

It is an advantage of this invention that the bile acid and thecarbohydrate remain in solution without precipitation at any pH fromacidic to alkaline. These aqueous solution systems of bile acid aresubstantially free of precipitate or particles. A further advantage ofthis invention is that the aqueous solution systems demonstrate nochanges in physical appearance such as changes in clarity, color or odorfollowing the addition of strong acids or alkali even after severalmonths observation under accelerated conditions of storage at 50° C.

In some embodiments of the invention, an aqueous solution system of bileacid is administered orally whereupon it reaches the intestine throughthe gastrointestinal track without precipitation of bile acids as solidsby exposure to acidic gastric juices and alkaline juices of theintestine. These dissolved bile acid formulations demonstrate intactsolution systems in the intestine can be effectively and completelyabsorbed and, consequently, undergo enterohepatic cycling. According tothe invention, bile acid solubility (e.g. precipitation and changes inphysical appearance) is unaffected by whether a carboxylic acid sidechain of certain bile acids can be protonated (non-ionized), ionized, ora simple carboxylic acid.

The ionization state of a bile acid carboxylic acid side chain greatlyeffects the hydrophobicity and the hydrophillicity of the bile acid inthese aqueous solution systems. In some embodiments of the invention,that ionization state is manipulated by adjusting the pH to control thetoxicity, absorption, and amphiphilicity of bile acids. One or more bileacid may be dissolved in these aqueous solution systems as atherapeutically active agent, as an adjuvant of a drug, as a carrier ofdrug or as an enhancer of drug solubility. These aqueous solutionsystems may be prepared for oral consumption, enemas, mouthwashes,gargles, nasal preparations, otic preparations, injections, douches,topical skin preparations, other topical preparations, and cosmeticpreparations which have a desired pH without the disadvantage ofprecipitation or deterioration in physical appearance after long periodsof time.

Soluble bile acids are any type of aqueous soluble bile acids. A bileacid salt is any aqueous soluble salt of a bile acid. The soluble bileacid derivatives of this invention are those derivatives which are assoluble or more soluble in aqueous solution than is the correspondingunderivatized bile acid. Bile acid derivatives include, but are notlimited to derivatives formed at the hydroxyl and carboxylic acid groupsof the bile acid with other functional groups including but not limitedto halogens and amino groups. Aqueous dissolved salts of bile acids maybe formed by the reaction of bile acids described above and an amineincluding but not limited to aliphatic free amines such as trientine,diethylene triamine, tetraethylene pentamine, and basic amino acids suchas arginine, lysine, omithine, and ammonia, and amino sugars such asD-glucamine, N-alkylglucamines, and quaternary ammonium derivatives suchas choline, heterocyclic amines such as piperazine, N-alkylpiperazine,piperidine, N-alkylpiperidine, morpholine, N-alkylmorphline,pyrrolidine, triethanolamine, and trimethanolamine. According to theinvention, aqueous soluble metal salts of bile acids, inclusion compoundbetween the bile acid and cyclodextrin and its derivatives, and aqueoussoluble 0-sulfonated bile acids are also included as soluble bile acidsalts. Soluble bile acid may include an aqueous preparation of a freeacid form of bile acid combined with one of HCl, acetic acid, ammonia,or arginine.

Bile acids used in this invention include, but are not limited toursodeoxycholic acid, chenodeoxycholic acid, cholic acid, hyodeoxycholicacid, deoxycholic acid, 7-oxolithocholic acid, lithocholic acid,iododeoxycholic acid, iocholic acid, tauroursodeoxycholic acid,taurochenodeoxycholic acid, taurodeoxycholic acid, taurolithocholicacid, glycoursodeoxycholic acid, taurocholic acid, glycocholic acid, andtheir derivatives at a hydroxyl or carboxylic acid group on the steroidnucleus.

A major advantage of the instant invention is that by delivery of bileacid in solution, it achieves higher in vivo levels of bile acids thanconventional preparations. Therefore, the therapeutic potential of bileacid may be more fully acheived than previous formulations. The in vivolevels of bile acids attainable with existing formulations in which bileis incompletely solubilized are lower and require administration oflarger amounts of bile acids. Since bile acid is completely dissolved inthe inventive formulations, higher in vivo levels of bile acid may beachieved, even though lower doses are administered.

In some embodiments of the invention, a plurality of bile acids are usedin a single formulation. Mixtures of two or more bile salts of differinghydrophobic activity may behave as a single bile salt of an intermediatehydrophobic activity. As a result, detergent properties and the toxicityof mixtures of two bile acids of differing hydrophobic activity oftenare intermediate between the individual components.

Carbohydrates suitable for use in the invention include aqueous solublestarch conversion products and aqueous soluble non-starchpolysaccharides. For purposes of the invention, aqueous soluble starchconversion products are defined as follows:

-   1. They may be obtained under various pH conditions from the partial    or incomplete hydrolysis of starch.-   2. Non-limiting examples include maltodextrin, dextrin, liquid    glucose, corn syrup solid (dried powder of liquid glucose), soluble    starch, and soluble starch, preferably maltodextrin or corn syrup    solid, most preferably corn syrup solid. Particularly preferred are    MALTRINS M200, a corn syrup solid, and MALTRIN® M700 a maltodextrin,    both of which are manufactured by GPC®, Grain Processing Corporation    of Muscatine, Iowa. For the purpose of this invention, the term    “corn syrup” includes both corn syrup and liquid glucose.-   3. If polymeric, the polymer has at least one reducing end and at    least one non-reducing end. The polymer may be linear or branched.-   4. The molecular weight is from about 100 mass units to over 10⁶    mass units. High molecular weight aqueous soluble starch conversion    products are those having a molecular weight over 10⁵.

For purposes of the invention, aqueous soluble non-starchpolysaccharides are defined as follows:

-   1. They may be obtained under various pH conditions by various    hydrolytic or synthetic mechanisms.-   2. Non-limiting examples include to dextran, guar gum, pectin,    indigestible soluble fiber.-   3. If polymeric, the polymer has at least one reducing end and at    least one non-reducing end. The polymer may be linear or branched.-   4. The molecular weight is from about 100 mass units to over 10⁶    mass units. Preferably the molecular weight is over 10⁵ mass units.

The amount of high molecular weight aqueous soluble starch conversionproduct and/or soluble non-starch polysaccharide used in the inventionis at least the amount needed to render the chosen bile acid saltsoluble in the concentration desired and in the pH range desired. Inpreferred embodiments of the invention, the approximate minimal quantityof maltodextrin required to prevent the precipitation of bile acids fromthe aqueous solution dosage forms of the invention is 5 g for every 0.2g of ursodeoxycholic acid, 25 g for every 1 g of ursodeoxycholic acid,and 50 g for every 2 g of ursodeoxycholic acid in 100 mL of water. Inpreferred embodiments of the invention, the approximate minimal quantityof maltodextrin is 30 g for every 200 mg of chenodeoxycholic acid, 12 gfor every 200 mg of 7-ketolithocholic acid, 10 g for every 200 mg ofcholic acid and 50 g for every 200 mg of deoxycholic acid. In preferredembodiments of the invention, the approximate minimal quantity of liquidglucose (commercial light corn syrup) required to prevent theprecipitation of bile acids from the aqueous solution dosage forms ofthe invention is 80 g for every 500 mg ursodeoxycholic acid in 100 mLwater, and 80 g for every 500 mg ursodeoxycholic acid in 200 mL water.In preferred embodiments of the invention, the approximate minimalquantity of dried powder of liquid glucose (corn syrup solid, e.g.,MALTRIN® M200) required to prevent the precipitation of bile acids fromthe aqueous solution dosage forms of the invention is 30 g for every 500mg ursodeoxycholic acid in 100 mL water, and approximately 30 g forevery 500 mg of ursodeoxycholic acid in 200 mL water. In preferredembodiments of the invention, the approximate minimal quantity ofsoluble non-starch polysaccharide (e.g., pectin, guar gum, gum arabic)required to prevent the precipitation of bile acids from the aqueoussolution dosage forms of the invention is 50 g guar gum for every 500 mgursodeoxycholic acid in 100 mL water and 80 g of pectin for every 500 mgof ursodeoxycholic acid in 100 mL water. The minimal required quantityof high molecular weight aqueous soluble starch conversion products orsoluble non-starch polysaccharide is primarily determined by theabsolute quantity of bile acids in the solution formulation rather thanthe concentration.

In some embodiments of the invention, a formulation may comprisecyclodextrin.

In some embodiments of the invention, the formulation further comprisesdietary fiber. Non-limiting examples of dietary fiber include guar gum,pectin, psyllium, oat gum, soybean fiber, oat bran, corn bran, celluloseand wheat bran.

In some embodiments of the invention, the formulation further comprisesemulsifying agents. For the purpose of the invention, the term“emulsifying agent” includes emulsifying agents and suspending agents.Non-limiting examples of emulsifying agents include guar gum, pectin,acacia, carrageenan, carboxymethyl cellulose sodium, hydroxymethylcellulose, hydroxypropyl cellulose, methyl cellulose, polyvinyl alcohol,povidone, tragacanth gum, xanthan gum, and sorbitan ester.

The selected pH range for which the formulation will not precipitate itsbile acid, starch conversion product, soluble non-starch polysaccharideor its pharmaceutical compound may be any range of pH levels obtainablewith an aqueous system. Preferably this range is between about pH 1 andabout pH 14 and more preferably between about pH 1 and about pH 10.Still more preferably the range is any subset of the range of pH levelsobtainable in an aqueous system sufficient for the pharmaceuticalformulation to remain in solution from preparation, to administration,to absorption in the body, according to the method of administration.

The invention contemplates the use of a broad range of pharmaceuticalcompounds. Non-limiting examples include hormones, hormone antagonists,analgesic, antipyretics, antiinflammatory drugs, immunoactive drugs,antineoplastic drugs, antibiotics, anti-inflammatory agents,sympathomimetic drugs, anti-infective drugs, anti-tumor agents, andanesthetics. Further non-limiting examples include drugs that target oreffect the gastrointestinal tract, liver, cardiovascular system, andrespiratory system. Further non-limiting examples of pharmaceuticalcompounds include insulin, heparin, calcitonin, ampicillin, octreotide,sildenafil citrate, calcitriol, dihydrotachysterol, ampomorphine,yohimbin, trazodone, acyclovir, amantadine-HCl, rimantadine-HCl,cidofovir, delavirdine-mesylate, didanosine, famciclovir, forscarmetsodium, fluorouracil, ganciclovir sodium, idoxuridine, interferon-α,lamivudine, nevirapine, penciclovir, ribavirin, stavudine, trifluridine,valacyclovir-HCl, zalcitabine, zidovudine, indinavir-H₂SO₄, ritonavir,nelfinavir-CH₃SO₃H, saquinavir-CH₃SO₃H, d-penicillamine, chloroquine,hydroxychloroquine, aurothioglucose, gold sodium thiomalate, auranofinlevamisole, sodium diethyldithiocarbamate, isoprinosine, methyl inosinemonophosphate, muramyl dipeptide, diazoxide, hydralazine-HCl, minoxidil,dipyridamole, isoxsuprine-HCl, niacin, nylidrin-HCl, phentolamine,doxazosin-CH₃SO₃H, prazosin-HCl, terazocin-HCl, clonidine-HCl,nifedipine, molsidomine, amiodarone, acetylsalicylic acid, verapamil,diltiazem, nisoldipine, isradipine, bepridil, isosorbide-dinitrate,pentaerythrytol-tetranitrate, nitroglycerin, cimetidine, famotidine,nizatidine, ranitidine, lansoprazole, omeprazole, misoprostol,sucralfate, metoclopramide-HCl, erythromycin, bismuth compound,alprostadil, albuterol, pirbuterol, terbutaline-H₂SO₄, salmetrol,aminophylline, dyphylline, ephedrine, ethylnorepinephrine, isoetharine,isoproterenol, metaproterenol, nedocromil, oxytriphylline, theophylline,bitolterol, fenoterol, budesonide, flunisolide,beclomethasone-dipropionate, fluticasone-propionate, codeine, codeinesulfate, codeine phosphate, dextromethorphan-HBr,triamcinolone-acetonide, montelukast sodium, zafirlukast, zileuton,cromolyn sodium, ipratropium bromide, nedocromil sodium benzonate,diphenhydramine-HCl, hydrocodone-bitartarate, methadone-HCl, morphinesulfate, acetylcysteine, guaifenesin, ammonium carbonate, ammoniumchloride, antimony potassium tartarate, glycerin, terpin-hydrate,colfosceril palmitate, atorvastatin-calcium, cervastatin-sodium,fluvastatin-sodium, lovastatin, pravastatin-sodium, simvastatin,picrorrhazia kurrva, andrographis paniculata, moringa oleifera, albizzialebeck, adhata vasica, curcuma longa, momordica charantia, gynmemasylvestre, terminalia arjuna, azadirachta indica, tinosporia cordifolia,metronidazole, amphotericin B, clotrimazole, fluconazole, haloprogin,ketoconazole, griseofulvin, itraconazole, terbinafin HCl,econazole-HNO3, miconazole, nystatin, oxiconazole-HNO₃,sulconazole-HNO₃, cetirizine-2HCl, dexamethasone, hydrocortisone,prednisolone, cortisone, catechin and its derivatives, glycyrrhizin,glycyrrhizic acid, betamethasone, ludrocortisone-acetate, flunisolide,fluticasone-propionate, methyl prednisolone, somatostatin, lispro,glucagon, proinsulin, insoluble insulins, acarbose, chlorpropamide,glipizide, glyburide, metformin-HCl, repaglinide, tolbutamide, aminoacid, colchicine, sulfinpyrazone, allopurinol, piroxicam, tolmetinsodium, indomethacin, ibuprofen, diflunisal, mefenamic acid, naproxen,and trientine.

Additional pharmaceutical compounds that may be included in theformulation are any compounds which remain soluble when added to theformulation. With an additional pharmaceutical compound in theformulation, a bile acid in solution may act as an adjuvant, carrier, orenhancer for the solubility of certain therapeutically active agents,including, but not limited to, insulin (pH 7.4-7.8), heparin (pH 5-7.5),calcitonin, ampicillin, amantadine, rimantadine, sildenafil, neomycinsulfate (pH 5-7.5), apomorphine, yohimbin, trazodone, ribavirin,paclitaxel and its derivatives, retinol, and tretinoin, which aresoluble and stable in acid and/or alkali and can be added as needed intothese aqueous solution dosage forms of certain concentrations of bileacids in this invention. Certain therapeutically active agents,including, but not limited to, metformin HCl (pH 5-7), ranitidine HCl,cimetidine, lamivudine, cetrizine 2HCl (pH 4-5), amantadine,rimantadine, sildenafil, apomorphine, yohimbine, trazodone, ribavirinand dexamethasone, hydrocortisone, prednisolone, triamcinolone,cortisone, niacin, taurine, vitamins, naturally occurring amino acids,catechin and its derivatives, glycyrrhizal extract and its mainconstituents such as glycyrrhizin and glycyrrhizic acid, water solublebismuth compounds (e.g., bismuth sodium tartrate), and which are solubleand stable in acid and/or alkali can be added as needed into theseaqueous solution dosage formulations containing ursodeoxycholic acid inthis invention.

According to the invention bismuth compounds comprise an aqueous solublereaction product between a bismuth ion and a chelator. Non-limitingexamples of such chelators include citric acid, tartaric acid, malicacid, lactic acid and eidetic acid and alkalies. Non-limiting examplesof bismuth compounds include an ammonium salt of a bismuth chelationcomplex, bismuth citrate, bismuth gallate, bismuth sulphate, bismuthsubnitrate, bismuth subsalicylate, tripotassium dicitrato bismuthate,and bismuth sodium tartrate.

The invention contemplates the use of pH adjustable agents. Non-limitingexamples include HCl, H₂SO₄, HNO₃, CH₃COOH, citric acid, malic acid,tartaric acid, lactic acid, phosphate, eidetic acid and alkalies.

In some embodiements of the invention, the formulations may be used totreat human and mammalian diseases. The invention contemplates treatinggastrointestinal disorders, liver diseases, gall stones, andhyperlipidemia. Non-limiting examples of liver diseases includealcohol-induced liver diseases and non-alcohol-induced liver diseases.Non-limiting examples of gastrointestinal disorders include chronicgastritis, reflux gastritis, and peptic ulcer disease. Non-limitingexamples of non-alcohol-induced liver diseases include primary biliarycirrhosis, acute and chronic hepatitis, primary sclerosing cholangitis,chronic active hepatitis, and excess accumulation of fat in the liver.The invention further contemplates treating viral, bacterial and fungaldiseases. In some embodiments of the invention, a formulation isadministered to treat and/or erradicate Helicobacter pylori infection.In some embodiments of the invention, a formulation is administered totreat and/or erradicate hepatitis C virus infection, influenza A,Influenza C, parainfluenza 1, sendai, rubella, and pseudorabies virus.In some embodiments of the invention, a formulation is administered totreat acute or chronic inflammatory diseases. Non-limiting examples ofinflammatory diseases include bronchitis, chronic pharyngitis, andchronic tonsillitis. In some embodiments of the invention, a formulationis administered to treat hypercholersterolemia.

In some embodiments of the invention, the formulation is modified suchthat it may be administered as a liquid, solid, powder or tablet. Insome embodiments of the invention, the formulation is comprised in asyrup, thick syrup or paste. A non-limiting example of a syrup is asolution of maltodextrin wherein the concentration of maltodextrin isless than 1.0 kg/L. A non-limiting example of a thick syrup is asolution of maltodextrin wherein the concentration of maltodextrin isbetween 1.0 kg/L and 1.2 kg/L inclusive. A non-limiting example of apaste is a solution of maltodextrin wherein the concentration ofmaltodextrin is greater than 1.2 kg/L.

EXAMPLES

The stability of dosage formulations of the invention were evaluated bymeasuring the concentration of the relevant bile acid over time inpreparations comprising soluble bile acid, a high molecular weightaqueous soluble starch conversion product, and water at various pH andtemperature levels. The retention time of each bile acid may be adjustedas needed to permit individual analysis each bile acid present in acomplex samples, i.e. a sample having a plurality of bile acids.

The stability tests were conducted on three different aqueous solutionsystems:

-   -   1. A bile acid and a high molecular weight aqueous soluble        starch conversion product were combined in aqueous solution        according to Example I, with results as shown in Tables 1A and        1B.    -   2. Mixed bile acids and high molecular weight aqueous soluble        starch conversion products were combined in aqueous solution        according to Example II, with results as shown in Table 2.    -   3. Bile acids, high molecular weight aqueous soluble starch        conversion products and branched chained amino acids (e.g.        leucine, isoleucine, valine, or other amino acid with a branched        side chain) were combined in aqueous solution according to        Example IV, with results as shown in Tables 3A through 3F.

The stability tests were performed with HPLC and microscope light atvarious pH conditions under the normal and accelerated conditions.Accelerated conditions for testing pharmaceutical compositions have beendescribed (Remington, The Science and Practice of Pharmacy, 19th ed., p.640). All of these stability test results were satisfactory in that theconcentration of bile acid as measured by HPLC did not changeappreciably over time at various pH levels. Thus, the formulations ofthe examples are suitable for preparing a commercial liquid dosage form.Particularly, all solution formulations which contained bile acid showedexcellent results in the stability tests with no precipitation and nophysical appearance changes over the test period. Some formulationsremain stable for over 2 years.

Moreover, the solution stability tests were conducted on the aqueoussolution dosage forms comprising the mixture of aqueous soluble UDCA,branched chained amino acid (leucine, isoleucine, valine) andmaltodextrin according to example IV as a typical example of thesolution dosage forms in which bile acid as a therapeutically activeagent, as an adjuvant or carrier, pharmaceutically active agent, orenhancer of solubility, and high molecular weight aqueous soluble starchconversion products or soluble non-starch polysaccharides are dissolved.According to the test results, there is no discoloration, no claritychanges, and no precipitation. Furthermore, there are no detectableimpurities from the deterioration of UDCA or branched chained aminoacids when examined by HPLC at various pH conditions such as pH 1, 3, 5,7, 9, and 10 under the accelerated conditions or incubation at (50° C.).

The aqueous solution dosage forms according to this invention did notchange either physically or chemically at various pH conditions underthe accelerated conditions despite the addition of therapeutically andchemically active agents that are stable and soluble in hydrochloricacid solution. Therefore, these aqueous solution systems are extremelyvaluable pharmaceutical dosage forms for the therapeutically active bileacids preparations, and/or the drug (pharmaceutical compound) deliverypreparations in which bile acids play roles as the adjuvant of drug, thecarrier of drug, or the enhancer of solubility of a drug by micelleformation at various pH conditions without the stability problems,including precipitation in acidic conditions.

For the solution stability test for each bile acid, HPLC was used tomeasure the concentration of the relevant soluble bile acid under thefollowing conditions: the elution solvent of 0.02 M KH₂PO₄:acetonitrilein a ratio of 55:45, with a pH of 3.01, the flow rate was 0.8 mL/min.,the injection volume was 20 μL, wave length for detection was 195 nm. Inthe tables, the concentration of the indicated bile acid salt for eachof the three numbered trials and the average thereof is reported on eachline. The percentage indicates the relative concentration of the bileacid salt after incubation for a certain amount of time in comparisonwith the initial concentration.

Example I

Solution dosage forms that were prepared according to the followingguidelines did not show any precipitation at any pH tested.

Soluble bile acid 200 mg (as free acid) Minimal quantity of maltodextrinfor CDCA: approx. 30 g; for UDCA: approx. 5 g; for 7-ketolithocholicacid: approx. 12 g; for cholic acid: approx. 10 g; for deoxycholic acid:approx. 50 g; for hyodeoxycholic acid: approx. 3.5 g Purified water tomake 100 mL

100 mL of the aqueous solution in which one of the above bile acids isdissolved was prepared. Into the resulting clear solution, maltodextrin,a high molecular weight aqueous soluble starch conversion product, wasadded with agitation at room temperature. Purified water was added toadjust the total volume to be 100 mL. According to the instant inventionand all examples, purified water is deionized, distilleddeionized-distilled water, or a grade commonly used for pharmaceuticalpreparations.

Based on these formulas, the aqueous solution dosage forms of variousconcentrations of certain bile acids (or salts) with its correspondingminimal quantity or more of high molecular weight aqueous soluble starchconversion products (for example; maltodextrin, liquid glucose, driedpowder of liquid glucose (commercial corn syrup solid), dextran,dextrin, and soluble starch) or soluble non-starch polysaccharide (e.g.guar gum, pectin, gum arabic) were prepared.

Example II

Solution dosage forms that were prepared according to the followingguidelines did not show any precipitation at any pH tested.

Soluble cholic acid 200 mg (as free acid), Soluble 7-ketolithocholicacid 200 mg (as free acid), Soluble chenodeoxycholic acid 200 mg (asfree acid), Minimal quantity of maltodextrin 40 g, and Purified water tomake 100 mL

60 mL of the aqueous solution in which soluble cholic acid, soluble7-ketolithocholic acid, and soluble chenodeoxycholic acid are dissolved,was prepared. Into the resulting clear solution, maltodextrin was addedwith agitating at room temperature. Purified water was added to adjustthe total volume to be 100 mL.

Using this formulation, the stability test for the aqueous solution ofthe mixture of various bile acids which can control the hydrophillicityor hydrophobicity was conducted.

Table 1A shows the results of a test of stability over time at pH 7 and50° C. of formulations of CA, 7-ketolithocholic acid, CDCA and DCA insolution with maltodextrin prepared according to Example I. Theconcentrations of the bile acids were measured by HPLC and theconcentration of the bile acid as a percentage of its concentration onday 0 is reported in the column labeled percentage.

Table 1B shows the results of the test of stability over time at pH 10and 50° C. of formulations of CA, 7-ketolithocholic acid, CDCA and DCAin solution with maltodextrin prepared according to Example I.

Table 2 shows results of the test of stability over time at pH 1 and 50°C. of formulations of CA, 7-ketolithocholic acid, CDCA and DCA insolution with maltodextrin at pH 1 and 50° C. prepared according toExample II.

TABLE 1A Day #1 #2 #3 Average Percentage CA 0 0.529 0.530 0.522 0.527100.0 4 0.460 0.524 0.524 0.502 95.4 7 0.520 0.525 0.547 0.531 100.8 200.516 0.576 0.535 0.542 103.0 KLCA 0 0.888 0.879 0.874 0.880 100.0 40.871 0.887 0.888 0.882 100.2 7 0.897 0.893 0.888 0.893 101.4 20 0.8930.909 0.894 0.899 102.1 CDCA 0 0.572 0.539 0.530 0.547 100.0 4 0.5400.552 0.576 0.556 101.6 7 0.581 0.588 0.553 0.574 105.0 20 0.565 0.6080.560 0.578 105.7 DCA 0 0.499 0.491 0.489 0.493 100.0 4 0.501 0.5000.474 0.491 99.6 7 0.488 0.487 0.484 0.486 98.6 20 0.478 0.476 0.4720.475 96.3

TABLE 1B Day #1 #2 #3 Average Percentage CA 0 0.534 0.524 0.490 0.516100.0 4 0.501 0.509 0.524 0.511 99.1 7 0.552 0.518 0.533 0.534 103.6 200.535 0.563 0.548 0.549 106.4 KLCA 0 0.879 0.874 0.857 0.870 100.0 40.870 0.873 0.880 0.874 100.5 7 0.893 0.876 0.882 0.884 101.5 20 0.8870.893 0.887 0.889 102.2 CDCA 0 0.541 0.532 0.495 0.522 100.0 4 0.5110.519 0.538 0.523 100.0 7 0.564 0.527 0.540 0.544 104.1 20 0.556 0.5690.558 0.561 107.4 DCA 0 0.491 0.488 0.471 0.483 100.0 4 0.493 0.4870.472 0.484 100.2 7 0.479 0.488 0.479 0.482 99.7 20 0.468 0.478 0.4790.475 98.3

TABLE 2 Day #1 #2 #3 Average Percentage CA 0 0.516 0.509 0.503 0.509100.0 4 0.453 0.453 0.466 0.457 89.8 7 0.434 0.426 0.468 0.443 86.9 200.207 — 0.206 0.207 40.6 KLCA 0 0.883 0.877 0.869 0.876 100.0 4 0.8700.866 0.847 0.861 98.3 7 0.848 0.844 0.843 0.845 96.4 20 0.661 — 0.6510.656 74.9 CDCA 0 0.560 0.528 0.513 0.534 100.0 4 0.488 0.510 0.5190.506 94.7 7 0.460 0.469 0.463 0.464 87.0 20 0.169 — 0.154 0.161 30.2

Example III

Solution dosage forms that were prepared according to the followingguidelines did not show any precipitation at any pH tested.

Soluble UDCA 200 mg (100 mg-2000 mg as free base) Minimal quantity ofmaltodextrin approx. 5 g (approx.1.25 g-50 g) Preservatives q.s.Flavoring agent q.s. Sweetener q.s. Purified water to 100 mL

80 mL of an aqueous solution in which soluble UDCA is dissolved wasprepared, and then, maltodextrin was added into the clear solution withagitating at room temperature. Into the resulting clear solution,sweetener, preservatives and flavoring agents were added in quantitiessuitable for a pharmaceutical formulation. Purified water was added toadjust the total volume to be 100 mL.

In these formulas, the aqueous solution dosage forms of variousconcentrations of ursodeoxycholic acid (or its salts) with itscorresponding minimal quantity or more of aqueous soluble starchconversion products (for example, maltodextrin, liquid glucose, driedpowder of liquid glucose (commercial corn syrup solid), dextran, orsoluble starch).

FIG. 6 is an NMR spectrum of UDCA illustrating that UDCA, when in acomposition prepared according to Example III, is absolutely free UDCA.That is, the carboxylic acid of UDCA at C-24 is the free form (R-COOH)and two hydroxy group at C-3 and C-7 are in the free form (R-OH)

In addition, the HPLC profile of UDCA in a composition preparedaccording to Example III (FIG. 7) is similar to the profile of of UDCAdissolved in methanol (FIG. 8). This data shows that there is noUDCA-complex compound. There is only free UDCA. The UDCA standardsolution was prepared by dissolving 100 mg UDCA in 100 mL of methanol. Amixture of acetonitrile (51), water (49), and acetic acid (1) was usedas the mobile phase.

Example IV

Solution dosage forms that were prepared according to the followingguidelines did not show any precipitation at any pH within the selected,desired range of pH values.

Soluble UDCA 0.2 g (1 g-2 g as free acid) Maltodextrin 5 g (35 g-50 g)Branched chained amino acid 15 g (5 g-15 g as free base) (e.g. leucine,isoleucine, valine) Sweetener q.s. Flavoring agent q.s. Purified waterto 100 mL

85 mL of the aqueous solution in which soluble UDCA is dissolved wasprepared, and then maltodextrin was added into the clear solution. Intothe resulting clear solution, branched amino acids were added withadjusting the pH (4-7) with agitation and then sweetener, preservatives,and flavoring agent were added. Purified water was added to adjust thetotal volume to be 100 mL.

Based on these formulations, the aqueous solution dosage forms ofvarious concentrations of ursodeoxycholic acid (or its salt) and itscorresponding minimal quantity or more of maltodextrin, liquid glucose,dried powder of liquid glucose (commercial corn syrup) or dextran) withvarious quantities of branched amino acid (total amount of leucine,isoleucine and valine) were prepared.

Tables 3A to 3F show stability test results over time of formulationprepared with amino acids according to Example IV. All stability testswere conducted at 50° C. Stability test results at pH 1 (Table 3A), pH 3(Table 3B), pH 5 (Table 3C), pH 7 (Table 3D), pH 9 (Table 3E), and pH 10(Table 3F) are shown.

TABLE 3A Stability of UDCA solution according to Example IV at pH1, 50°C. Day #1 #2 #3 Average Percentage Ile 0 0.261 0.236 0.249 0.248 100.0 10.256 0.275 0.251 0.261 105.0 2 0.268 0.263 0.251 0.260 104.9 6 0.2950.268 0.291 0.285 114.6 7 0.249 0.254 0.267 0.257 103.4 8 0.253 0.2430.240 0.245 98.8 9 0.263 0.268 0.263 0.265 106.6 Leu 0 0.485 0.428 0.4700.461 100.0 1 0.470 0.477 0.456 0.468 101.5 2 0.485 0.481 0.460 0.475103.1 6 0.553 0.510 0.529 0.531 115.1 7 0.478 0.473 0.513 0.488 105.8 80.474 0.454 0.511 0.480 104.0 9 0.483 0.485 0.476 0.481 104.4 Val 00.506 0.448 0.460 0.471 100.0 1 0.438 0.458 0.471 0.456 96.7 2 0.4790.485 0.513 0.492 104.5 6 0.505 0.536 0.549 0.530 112.4 7 0.494 0.4650.496 0.485 102.9 8 0.488 0.491 0.459 0.479 101.7 9 0.479 0.496 0.4900.488 103.6 Sol 0 0.319 0.315 0.322 0.319 100.0 1 0.332 0.344 0.3510.342 107.4 2 0.371 0.339 0.403 0.371 116.4 6 0.396 0.409 0.411 0.405127.2 7 0.365 0.351 0.381 0.366 114.7 8 0.409 0.365 0.331 0.368 115.6 90.338 0.391 0.374 0.368 115.4 UDCA 0 0.388 0.387 0.389 0.388 100.0 10.367 0.370 0.366 0.368 94.8 2 0.374 0.388 0.388 0.383 98.9 6 0.3710.380 0.382 0.377 97.3 7 0.378 0.376 0.379 0.378 97.4 8 0.374 0.3820.384 0.380 97.9 9 0.370 0.367 0.370 0.369 95.1

TABLE 3B Stability of UDCA solution according to Example IV at pH 3, 50°C. Day #1 #2 #3 Average Percentage Ile 0 0.261 0.254 0.253 0.256 100.0 10.266 0.268 0.261 0.265 103.3 2 0.273 0.243 0.247 0.254 99.3 6 0.2960.306 0.300 0.301 117.4 7 0.247 0.265 0.257 0.256 100.0 8 0.250 0.2470.247 0.248 96.7 13 0.285 0.240 0.250 0.258 100.9 Leu 0 0.495 0.4650.452 0.471 100.0 1 0.489 0.480 0.470 0.480 101.9 2 0.495 0.472 0.4810.483 102.6 6 0.522 0.532 0.556 0.537 114.0 7 0.492 0.482 0.491 0.488103.7 8 0.543 0.515 0.495 0.517 109.9 13 0.512 0.496 0.543 0.517 109.8Val 0 0.485 0.491 0.498 0.491 100.0 1 0.467 0.481 0.446 0.465 94.6 20.510 0.493 0.527 0.510 103.8 6 0.527 0.491 0.553 0.524 106.6 7 0.4850.481 0.468 0.478 97.3 8 0.490 0.491 0.544 0.508 103.5 13 0.519 0.4980.517 0.511 104.1 Sol 0 0.343 0.355 0.370 0.356 100.0 1 0.340 0.3500.316 0.335 94.2 2 0.383 0.371 0.400 0.385 108.0 6 0.378 0.341 0.4160.378 106.3 7 0.355 0.381 0.315 0.350 98.4 8 0.343 0.350 0.395 0.363101.9 13 0.377 0.382 0.423 0.394 110.7 UDCA 0 0.395 0.396 0.393 0.395100.0 1 0.396 0.401 0.392 0.396 100.4 2 0.427 0.421 0.416 0.421 106.8 60.407 0.408 0.402 0.405 102.7 7 0.412 0.409 0.411 0.411 104.1 8 0.4150.418 0.408 0.414 104.9 13 0.415 0.412 0.416 0.414 105.0

TABLE 3C Stability of UDCA solution according to Example IV at pH 5, 50°C. Day #1 #2 #3 Average Percentage Ile 0 0.285 0.258 0.295 0.279 100.0 30.280 0.275 0.275 0.277 99.0 6 0.285 0.273 0.270 0.276 98.7 10 0.2740.276 0.276 0.275 98.4 13 0.273 0.287 0.278 0.279 100.0 17 0.278 0.2760.270 0.275 98.3 20 0.261 0.275 0.261 0.266 95.0 24 0.267 0.274 0.2920.277 99.3 Leu 0 0.495 0.467 0.535 0.499 100.0 3 0.510 0.495 0.494 0.500100.1 6 0.489 0.479 0.484 0.484 97.0 10 0.486 0.490 0.499 0.492 98.5 130.492 0.509 0.508 0.503 100.8 17 0.514 0.508 0.504 0.509 100.9 20 0.4990.500 0.499 0.499 101.1 24 0.488 0.509 0.528 0.508 101.9 Val 0 0.4830.498 0.481 0.487 100.0 3 0.492 0.494 0.526 0.504 103.4 6 0.459 0.4750.481 0.472 96.8 10 0.500 0.436 0.480 0.472 96.9 13 0.464 0.451 0.4740.463 95.0 17 0.407 0.491 0.462 0.453 93.0 20 0.471 0.512 0.477 0.48799.9 24 0.471 0.476 0.458 0.468 96.1 Sol 0 0.341 0.351 0.360 0.351 100.03 0.342 0.386 0.371 0.366 104.5 6 0.316 0.321 0.342 0.326 93.1 10 0.3410.299 0.335 0.325 92.7 13 0.355 0.326 0.350 0.344 98.0 17 0.334 0.3760.353 0.354 101.0 20 0.347 0.398 0.394 0.380 108.3 24 0.416 0.353 0.3780.382 109.0 UDCA 0 0.407 0.404 0.404 0.405 100.0 3 0.409 0.402 0.4030.405 99.9 6 0.410 0.403 0.409 0.407 100.6 10 0.404 0.405 0.407 0.405100.1 13 0.408 0.403 0.395 0.402 99.3 17 0.411 0.402 0.404 0.406 100.220 0.405 0.394 0.396 0.398 98.4 24 0.399 0.408 0.406 0.404 99.9

TABLE 3D Stability of UDCA solution according to Example IV at pH 7, 50°C. Day #1 #2 #3 Average Percentage Ile 0 0.296 0.289 0.281 0.289 100.0 50.300 0.282 0.281 0.288 99.7 8 0.277 0.282 0.268 0.276 95.5 12 0.2730.278 0.278 0.277 95.8 15 0.271 0.273 0.266 0.270 93.5 19 0.294 0.2850.281 0.287 99.3 Leu 0 0.519 0.513 0.495 0.509 100.0 5 0.499 0.499 0.4980.498 97.9 8 0.498 0.513 0.480 0.497 97.7 12 0.508 0.516 0.515 0.513100.9 15 0.503 0.505 0.499 0.502 98.7 19 0.521 0.509 0.516 0.515 101.3Val 0 0.483 0.530 0.525 0.513 100.0 5 0.502 0.447 0.499 0.483 94.1 80.488 0.498 0.493 0.493 96.2 12 0.490 0.469 0.443 0.467 91.2 15 0.4920.541 0.442 0.492 95.9 19 0.458 0.500 0.482 0.480 93.6 Sol 0 0.333 0.3520.363 0.349 100.0 5 0.344 0.309 0.349 0.334 95.6 8 0.334 0.379 0.3770.363 104.0 12 0.345 0.344 0.317 0.335 96.0 15 0.286 0.406 0.321 0.33896.7 19 0.338 0.416 0.351 0.368 105.4 UDCA 0 0.427 0.416 0.428 0.424100.0 5 0.406 0.427 0.432 0.422 99.4 8 0.419 0.408 0.417 0.414 97.7 120.414 0.418 0.419 0.417 98.4 15 0.413 0.418 0.409 0.414 97.5 19 0.4290.421 0.424 0.425 100.1

TABLE 3E Stability of UDCA solution according to Example IV at pH 9, 50°C. Day #1 #2 #3 Average Percentage Ile  0 0.291 0.286 0.282 0.286 100.0 3 0.266 0.273 0.282 0.273 95.6  6 0.277 0.274 0.272 0.274 95.9 10 0.2430.245 0.295 0.261 91.2 13 0.246 0.269 0.236 0.250 87.4 17 0.275 0.2800.245 0.267 93.1 Leu  0 0.509 0.513 0.511 0.511 100.0  3 0.485 0.4870.492 0.488 95.5  6 0.495 0.496 0.492 0.494 96.8 10 0.470 0.467 0.5280.488 95.6 13 0.461 0.491 0.450 0.467 91.5 17 0.468 0.516 0.500 0.49596.9 Val  0 0.508 0.476 0.484 0.489 100.0  3 0.463 0.487 0.485 0.47897.8  6 0.493 0.473 0.495 0.487 99.5 10 0.441 0.428 0.471 0.447 91.3 130.467 0.483 0.537 0.496 101.3 17 0.499 0.495 0.501 0.498 101.8 Sol  00.341 0.316 0.328 0.328 100.0  3 0.297 0.317 0.317 0.310 94.5  6 0.3130.291 0.314 0.306 93.2 10 0.268 0.253 0.324 0.282 85.8 13 0.270 0.2660.334 0.290 88.3 17 0.337 0.329 0.317 0.328 99.8 UDCA  0 0.389 0.3850.389 0.388 100.0  3 0.405 0.400 0.394 0.400 103.2  6 0.427 0.411 0.4160.418 107.9 10 0.420 0.418 0.450 0.429 110.8 13 0.465 0.434 0.441 0.447115.3 17 0.454 0.457 0.413 0.441 113.9

TABLE 3F Stability of UDCA solution according to Example IV at pH 10,50° C. Day #1 #2 #3 Average Percentage Ile 0 0.292 0.282 0.287 0.287100.0 2 0.253 0.237 0.239 0.243 84.7 5 0.221 0.212 0.221 0.218 76.0 70.219 0.215 0.207 0.214 74.5 9 0.206 0.192 0.207 0.202 70.2 Leu 0 0.5070.495 0.509 0.504 100.0 2 0.462 0.442 0.442 0.449 89.1 5 0.429 0.4280.427 0.428 85.0 7 0.410 0.417 0.414 0.414 82.1 9 0.417 0.377 0.4180.404 80.2 Val 0 0.480 0.506 0.471 0.486 100.0 2 0.536 0.478 0.504 0.506104.2 5 0.371 0.445 0.400 0.405 83.5 7 0.384 0.384 0.424 0.397 81.8 90.389 0.354 0.362 0.368 75.8 Sol 0 0.368 0.376 0.331 0.358 100.0 2 0.2840.257 0.266 0.269 75.1 5 0.053 0.217 0.192 0.154 43.0 7 0.042 0.0260.156 0.075 20.8 9 0.033 0.019 0.023 0.025 7.0 UDCA 0 0.416 0.402 0.4060.408 100.0 2 0.402 0.397 0.400 0.399 97.9 5 0.425 0.413 0.423 0.420103.0 7 0.406 0.402 0.408 0.406 99.4 9 0.424 0.426 0.421 0.423 103.8

Example V

Solution dosage forms that were prepared according to the followingguidelines did not show any precipitation at any pH within the selected,desired pH range. This formulation is based on the known analytical datafor pharmaceutical use of bear bile.

Tauro UDCA 7 g Tauro CDCA 1 g Glyco UDCA 0.8 g Glyco CDCA 0.2 g SolubleUDCA 1 g (or 3 g as free form) Aqueous soluble starch conversion product250 g. Sweetener q.s. Flavoring agent q.s. Purified water to 2.0 L

Soluble UDCA is dissolved in water and then high molecular weightaqueous soluble starch conversion product and water are added. Into theresulting clear solution, Tauro UDCA, Tauro CDCA, Glyco UDCA, GlycoCDCA, sweetener, and flavoring agent were added. Purified water wasadded to adjust the total volume to be 2.0 L.

Example VI

The aqueous solution dosage forms, according to this invention,containing 200 mg of ursodeoxycholic acid (UDCA), were preparedaccording to the method described in the above-described Example III andwere administered to three healthy men having normal body weight afterfasting. The hematic levels of UDCA and glyco UDCA were evaluated bymeans of well known chemical methods. After applying buffered serum tosep-pak column, methanol eluate was derivatized with phenacyl bromide at80° C. for 45 minutes. These phenacyl bromide derivatives were dissolvedin acetonitrile in preparation for HPLC. The experimental results of theabsorption measured at certain times after dosage administration includethe total absorption expressed as the area under the serumconcentration-time curve (AUC: jig/mL x hours), the maximum hematicconcentration (C_(max); μg/mL) that has been obtained, and the time(T_(max); hour) in which said maximum concentration has been obtained.These results are reported in Table 4, FIG. 1, and FIG. 2.

The experimental pharmacokinetic tests of the aqueous solution dosageforms according to this invention carried out on men show substantialimprovement in AUC, C_(max) and T_(max) in comparison with the bestresults from any dosage forms known presently. The maximum hematicconcentration (C_(max)) in Table 4 shows an average of 8.43±1.69 gg/mLwhich is at least two times higher than that reported for use of entericcoated sodium salt of UDCA preparations and four times higher than thatobtained using regular UDCA tablet preparations. Moreover, the time ofpeak concentration (T_(max)) which is related closely to the rate ofabsorption of UDCA from the aqueous solution dosage forms is 0.25 hours,at least three times faster than the fastest T_(max) previously known.

Table 4A and Table 4B show plasma concentration of UDCA and GUDCAmeasured in 3 men over time following on oral administration of the UDCAand GUL)CA containing formulations according to Example VI andcomparison of results against results of others employing differentpharmaceutical formulations of UDCA.

Table 5 shows phamacokinetic parameters of UDCA in human after an oraladministration of liquid formulation of UDCA. C_(max) is shown.

Taken together, the data in Tables 4 and 5 and FIGS. 3 and 4 illustratethe supperiority of formulations of the instant invention overconventional formulations with respect to C_(max) and T_(max). theinstant The inventive solutions were effect without any break-down ofthe solution system caused by the pH of the environment in the stomachand intestines. The therapeutic potential of bile acid and possibly evenadded pharmaceuticals may be more fully realized using the forulationsof the invention. When the therapeutically active ingredients in aqueoussolution forms are not precipitated as solid by acidic gastric juices inthe stomach and by the various alkaline pH levels of the intestine, theformulation overcomes as a natural consequence, the scarcebioavailability resulted by the unexpected, undesirable results for theextent and the rate of release by disintegration, dissolution and/ordiffusion should be overcome.

TABLE 4A Plasma concentration of UDCA and GUDCA after an oraladministration of this invention at a dose of 200 mg to three men UDCAGUDCA Time (h) #1 #2 #3 mean #1 #2 #3 mean 0.25 5.1202 10.9171 9.1598.43 ± 1.69 0.1419 0.4549 0.3328 0.31 ± 0.09 0.5 4.4528 7.7432 7.43956.55 ± 1.05 0.2564 1.2455 0.864 0.79 ± 0.29 1 1.6921 1.546 0.2163 1.15 ±0.47 0.2162 0.6926 0.2142 0.37 ± 0.16 1.5 0.5256 0.2759 0.168 0.32 ±0.11 1.1573 0.1929 0.4752 0.61 ± 0.29 2 0.2349 0.2176 0.1227 0.19 ± 0.030.4013 0.0312 0.0657 0.17 ± 0.12 3 0.1237 N.D. 0.2074 0.17 ± 0.04 0.50850.4303 0.3315 0.42 ± 0.05 5 1.9205 0.0229 1.6311 1.18 ± 0.61 7 0.53280.4797 0.91 0.64 ± 0.14 AUC (μg · h/mL) 4.32 6.6 5.47 5.46 ± 0.66 6.262.22 4.65 4.38 ± 1.17 C_(max) (μg/mL) 5.21 10.92 9.16 8.43 ± 1.69 1.921.25 1.63 1.6 T_(max) (h) 0.25 0.25 0.25 0.25 5 0.5 5 3.5 ± 1.5

TABLE 4B Pharmacokinetic parameters of UDCA in human after an oraladministration of UDCA (M ± S.E.) C_(max) (ug/mL) T_(max) (hr) Roda etal. (1994) UDCA gelatine capsule, 450 mg 2.59 3.8 NaUDC gelatinecapsule, 475 mg 3.42 2.4 NaUDC enteric-coated, 475 mg 10 3.4 Nagamatsuet al. (1997) UDCA 200 mg  1.9 ± 0.25 1.5 ± 0.4 UDCA 400 mg 7.09 ± 1.430.8 ± 0.2 UDCA in this invention, 200 mg 8.43 ± 1.69  0.25

TABLE 5A Pharmacokinetic parameter (C_(max)) of UDCA in human after oraladministration of a liquid solution containing 600 mg UDCA per day. TimePerson Person Person Person Person (min) #1 #2 #3 #4 #5 Average Std Dev 0 0.35 1.63 0.40 0.00 0.71 0.618 0.619  5 2.51 9.79 1.68 2.65 6.264.578 3.405 15 12.50 47.46 8.34 11.84 21.83 20.394 15.933 60 9.72 6.467.77 9.81 17.25 10.202 4.183 120  3.77 1.71 1.40 1.15 2.81 2.168 1.097240  0.65 0.93 0.50 0.48 1.30 0.772 0.346

TABLE 5B Pharmacokinetic parameter (C_(max)) of UDCA in human after anoral administration of a syrup containing 600 mg UDCA per day. TimePerson Person Person Person Person (min) #1 #2 #3 #4 #5 Average Std Dev 0 0.62 0.58 0.38 0.00 0.41 0.398 0.246  5 2.76 2.63 0.83 1.42 2.241.976 0.827 15 7.80 4.45 3.54 5.85 14.08 7.144 4.197 60 16.08 20.33 8.7612.06 17.77 15.000 4.605 120  3.98 4.24 5.09 7.79 3.00 4.820 1.820 240 0.81 0.99 1.47 1.85 1.17 1.258 0.411

Example VII

Solution dosage forms that were prepared according to the followingguidelines did not show any precipitation at any pH within the selecteddesired range of pH values.

Soluble UDCA 0.2 g(0.05 g-2 g as free acid) Dried powder of liquidglucose 20 g (3 g-120 g) (Commercial corn syrup solid) Soluble nonstarch polysaccharide 0.01 g(0.001 g-0.05 g) (Guar gum or pectin, etc.)Purified water to make 100 mL

85 mL of the aqueous solution in which soluble UDCA is dissolved wasprepared, and then the mixture of dried powder of liquid glucose, a highmolecular weight aqueous soluble starch conversion product and a solublenon starch polysaccharide (guar gum, pectin, etc.) was added into theclear solution. Purified water was added to adjust the total volume to100 mL.

Example VIII: Mixture Solution

The formulations of Examples VIII, IX, X, XI, and XII include bismuthsulfate. In each of these examples, solution dosage forms were preparedby adding an amount of an ammonium salt of bismuth sulfate sufficient toprovide the indicated amount of bismuth sulfate.

Solution dosage forms that were prepared according to the followingguidelines did not show any precipitation at any pH within the selecteddesired range of pH values.

UDCA 5 g CDCA 5 g Bismuth sulfate 5 g Corn syrup solid 260 g Citric acidq.s. Purified water to make 1.0 L

The UDCA and CDCA were first dissolved in 1.5 mL of a 1N NaOH solution.Next, to the resulting clear solution were added the bismuth citrate and150 mL of water. Then, the corn syrup solid was added portion by portionwith vigorous agitation. The resulting solution was titrated to pH 4with citric acid. Purified water was added to adjust the total volume to1.0 L.

Example IX: UDCA-Syrup (20 g UDCA/L)

Solution dosage forms that were prepared according to the followingguidelines did not show any precipitation at any pH within the selecteddesired range of pH values.

UDCA 20 g 1 N NaOH 60 mL Maltodextrin 700 g Bismuth sulfate 4 g Citricacid or lactic acid q.s. Purified water to make 1.0 L

The UDCA is first dissolved in 60 mL of a iN NaOH solution. Next, to theresulting clear solution were added the bismuth sulfate and 150 mrL ofwater. Then, the maltodextrin was added portion by portion with vigorousagitation. The resulting solution was titrated to pH 3.5 with citricacid. Purified water was added to adjust the total volume to 1.0 L.

Example X: UDCA-Syrup (20 g UDCA/L)

Solution dosage forms that were prepared according to the followingguidelines did not show any precipitation at any pH within the selecteddesired range of pH values.

UDCA 20 g 1 N NaOH 60 mL Corn syrup solid 1,050 g Bismuth sulfate 4 gCitric acid or lactic acid q.s. Purified water to make 1 L

The UDCA is first dissolved in 60 mL of a IN NaOH solution. Next, to theresulting clear solution were added the bismuth sulfate and 280 mL ofwater. Then, 1,050 g of corn syrup solid was added portion by portionwith vigorous agitation. The resulting solution was titrated to pH 3.5with citric acid. Purified water was added to adjust the total volume to1.0 L.

Example XI: UDCA-Thick Syrup (30 g UDCA/L)

Solution dosage forms that were prepared according to the followingguidelines did not show any precipitation at any pH within the selecteddesired range of pH values.

UDCA 30 g 1 N NaOH 90 mL Maltodextrin 1,050 g Citric acid or lactic acid50 g Purified water to make 1.0 L

The UDCA is first dissolved in 90 mL of a iN NaOH solution. Next, to theresulting clear solution were added the bismuth sulfate and 250 mL ofwater. Then, 1,050 g of maltodextrin was added portion by portion withvigorous agitation. The resulting solution was titrated to pH 3 by theaddition of 50 g of citric acid. Purified water was added to adjust thetotal volume to 1.0 L.

Example XII: UDCA-Thick Syrup (30 g UDCA/L)

Solution dosage forms that were prepared according to the followingguidelines did not show any precipitation at any pH within the selecteddesired range of pH values.

UDCA 30 g 1 N NaOH 90 mL Corn syrup solid 1,500 g Citric acid or lacticacid 50 g Purified water to make 1.0 L

The UDCA is first dissolved in 90 mL of a iN NaOH solution. Next, to theresulting clear solution were added the bismuth sulfate and 230 mL ofwater. Then, 1,500 g of corn syrup solid was added portion by portionwith vigorous agitation. The resulting solution was titrated to pH 3 bythe addition of 50 g of citric acid. Purified water was added to adjustthe total volume to 1.0 L.

Example XIII: UDCA-Paste (45 g UDCA/L)

The formulations of Examples XIII, XIV, XV, XVI, and XVIII includebismuth citrate. In each of these examples, solution dosage forms wereprepared by adding an amount of an ammonium salt of bismuth citratesufficient to provide the indicated amount of bismuth citrate.

Solution dosage forms that were prepared according to the followingguidelines did not show any precipitation at any pH within the selecteddesired range of pH values.

UDCA 45 g 1 N NaOH 135 mL Maltodextrin 1,575 g Bismuth citrate 10 gCitric acid or lactic acid q.s. Purified water to make 1.0 L

The UDCA is first dissolved in 135 mL of a 1 N NaOH solution. Next, tothe resulting clear solution were added the bismuth citrate and 200 mLof water. Then, 1,575 g of maltodextrin was added portion by portionwith vigorous agitation. The resulting solution was titrated to pH 3 bythe addition of citric acid. Purified water was added to adjust thetotal volume to 1.0 L.

Five human subjects were provided with dosage forms prepared accordingto this Example. The results are shown in Tables 5A and 5B and renderedgraphically in FIGS. 3 and 4. A comparrison of the sharp peak of FIG. 3with the broad peak of FIG. 4 indicates that, by adjusting the dosageform, a practitioner may manipulate the bile acid C_(max) and T_(max).

H. pylori were cultured on Columbia Bood Agar Base (CRAB) mediacontaining a preparation of Example IX. 2 L of CRAB plates were preparedwhich contained 9.9 g of CRAB, 9.1 g of tryptic soy agar, 50 mL ofsheeps blood, vacomycin, amphotericin B, polymixin B, 2 mL of ExampleIX, and 358 mL distilled water. After 48 or 72 hours of microaerophillicincubation, bateria were fixed using Kamovsky's fixative and embedded inepon. Electron micrographs of H. pylori cells are shown in figures FIGS.5A to 5C.

Example XIV: UDCA-Paste (45 g UDCA/L)

Solution dosage forms that were prepared according to the followingguidelines did not show any precipitation at any pH within the selecteddesired range of pH values.

UDCA 45 g 1 N NaOH 135 mL Corn syrup solid 2,300 g Citric acid or lacticacid 50 g Purified water to make 1.0 L

The UDCA is first dissolved in 135 mL of a iN NaOH solution. Next, tothe resulting clear solution were added the bismuth citrate and 150 mLof water. Then, 2,300 g of corn syrup solid was added portion by portionwith vigorous agitation. The resulting solution was titrated to pH 3 bythe addition of citric acid. Purified water was added to adjust thetotal volume to 1.0 L.

Example XV: Mixture solution of UDCA (22 g) and CDCA (3 g)

Solution dosage forms that were prepared according to the followingguidelines did not show any precipitation at any pH within the selecteddesired range of pH values.

UDCA 22 g 1 N NaOH 75 mL CDCA 3 g Maltodextrin 875 g Bismuth citrate 4 gCitric acid or lactic acid q.s. Purified water to make 1.0 L

The UDCA and CDCA are first dissolved in 75 mL of a 1N NaOH solution.Next, to the resulting clear solution were added the bismuth citrate and240 mL of water. Then, 875 g of maltodextrin was added portion byportion with vigorous agitation. The resulting solution was titrated topH 3 by the addition of citric acid. Purified water was added to adjustthe total volume to 1.0 L.

Example XVI: Mixture solution of UDCA (22 g) and CDCA (3 g)

Solution dosage forms that were prepared according to the followingguidelines did not show any precipitation at any pH within the selecteddesired range of pH values.

UDCA 22 g 1 N NaOH 75 mL CDCA 3 g Corn syrup solid 1,320 g Bismuthcitrate 4 g Citric acid or lactic acid q.s. Purified water to make 1.0 L

The UDCA and CDCA are first dissolved in 75 mL of a 1 N NaOH solution.Next, to the resulting clear solution were added the bismuth citrate and240 mL of water. Then, 1,320 g of corn syrup solid was added portion byportion with vigorous agitation. The resulting solution was titrated topH 3 by the addition of citric acid. Purified water was added to adjustthe total volume to 1.0 L.

Example XVII

The effect of treating H. pylori infected mice with a solution dosageform of the invention was tested. Six week old C57BL/6 female mice wereinfected by feeding at diet comprising 10⁹ CFU/mL H. pylori, SS1 strain.The animals consumed this feed twice, one week apart. Subsequently, 0.2mL of a solution dosage form according to Example VIII was administeredto four infected animals once per day for one week. Two animals weresacrificed one week following administration of the last dose of theinventive solution. The remaining two animals were sacrificed four weeksfollowing administration of the last dose of the inventive solution.Whole stomachs were washed with saline to remove mucosa and debris. Asample of stomach tissue from each animal was subjected to a CLO testusing a rapid urease test kit (Delta West, Australia). Each residualstomach was fixed with 10% formalin solution and embedded with paraffin.Sections (4 μm thick) were collected on glass slides and stained withH&E staining solution and Warthin staining solution. Tissue wasevaluated for pathological status by conventional light microscopy.

The results, summarized in Table 6, indicate that the urease testresults were negative for mice passed one week after discontinuingadministration of the liquid dosage form, and H. pylori was not seen inWarthin examination. Of the other two mice, one showed a negative ureasetest and no H. pylori were seen by Warthin examination. The other,however, yielded a positive urease test although only a few H. pyloriwere seen in Warthin examination.

TABLE 6 Weeks After Warthin Treatment Animal Urease Test Examination 1 1Negative No H. pylori 1 2 Negative No H. pylori 4 3 Negative No H.pylori 4 4 Positive A few H. pylori

Example XVIII

Assays for growth of H. pylori on media containing UDCA, bismuth citrateor both UDCA and bismuth citrate were performed. For these assays thefollowing media was used:

-   -   000112B-1 having a pH of 4.0 and comprising 525 g/L maltodextrin        and 15 g/L UDCA.    -   OSABY having a pH of 3.7 and comprising 1 kg/L corn syrup solid        and 6 g/L bismuth citrate.

Three assays were performed to assess the growth capacity of H. pyloriin the presence of UDCA, bismuth or both wherein the pH, concentration,and length of exposure was varied.

-   1. Helicobacter pylori was suspended in physiological saline to give    about 10⁹ organisms per milliliter. 50 μL of this innoculum was    transferred to tubes containing 1 mL of citrate-phosphate buffer at    pH 3.0, 4.0, and 4.5. Paired tubes were prepared with and without 6    mM Urea. Following a 30 minute room temperature incubation, the    suspensions were subcultured on agar plates containing 000112B-1    using a 1 μL loop. Plates were incubated microaerophilically at    37° C. for 72 hours. This procedure is illustrated in FIG. 9.

As shown in Table 7, H. pylori grew poorly on pH 3 and pH 4 controlmedia. Table 7 further shows that H. pylori does not grow on pH 3 and pH4 media containing UDCA. The designations “3 ml”, “4 ml” and “5 ml”refer to the total volume of 000112B-1 media per plate. “PBS” isphosphate buffered saline at pH 7.0.

TABLE 7 Urease Test Plate pH Urea 1-2 sec. 10 min. 2 hr. 20 hr. Control3.0 Yes FO FO O O No FO FO O O 4.0 Yes FO FO O O No FO FO O P 4.5 Yes FPFP P P No FO O FP P PBS Yes O FP P P No O FP P P 000112B-1 3.0 Yes Y Y YY (3 mL) No Y Y Y Y 4.0 Yes Y Y FO O No Y Y Y FO 4.5 Yes FP FP P P No FPFP P P 000112B-1 3.0 Yes Y Y Y Y (4 mL) No Y Y Y Y 4.0 Yes Y Y Y FO No YY FO FO 4.5 Yes FP FP P P No FP FP FP P 000112B-1 3.0 Yes Y Y Y Y (5 mL)No Y Y Y Y 4.0 Yes Y Y FO FO No Y Y Y Y 4.5 Yes FP FP P P No FP FP P PKey Y FO O FP P Color Yellow Faint Orange Orange Faint Pink PinkHelicobacter None Very Rare Rare Exist Many

-   2. Helicobacter pylori was suspended in physiological saline to give    about 10⁹ organisms per milliliter. 50 μL of this innoculum was    transferred to tubes containing 1 mL of citrate-phosphate buffer at    various concentrations of plating media such as 1/10, 1/30, 1/50,    1/100, 1/200, 1/500, 1/800, 1/1000, 1/2000. All tubes were prepared    with 6 mM Urea. Following a 30 minute room temperature incubation,    the suspensions were subcultured on agar plates using a 1 μL loop.    These plates were substantially free of bismuth and bile acids.    Plates were incubated microaerophilically at 37° C. for 72 hours.    This procedure is illustrated in FIG. 10.

Table 8 shows urease test results following 72 hours of growth of H.pylori on media prepared with dilutions of UDCA (000112B-1) bismuthcitrate (OSABY) or both UDCA and bismuth citrate. Poor growth of H.pylori on media containing either UDCA or bismuth citrate was observed(Table 8). Growth of H. pylori was fuirther attenuated when cultured onmedia containing both UDCA and bismuth citrate (Table 8).

TABLE 8 Urease Test Plate Immediately 10 min. 30 min. 60 min. 000112B-1Control P P P P 1/10 Y FP P P 1/30 Y FP P P 1/50 Y FP P P 1/100 Y FP P P1/200 Y FP P P 1/500 Y FP P P 1/800 FP P P P 1/1000 FP P P P 1/2000 FP PP P OSABY Control P P P P 1/10 Y FP P P 1/30 Y FP P P 1/50 Y FP P P1/100 FP FP P P 1/200 FP FP P P 1/500 FP FP P P 1/800 FP P P P 1/1000 FPP P P 1/2000 FP P P P 000122B-1 + OSABY Control P P P P 1/10 Y FP FP FP1/50 Y Y Y Y 1/100 Y Y Y Y 1/500 Y FP FP FP 1/1000 Y FP P P Key Y FO OFP P Color Yellow Faint Orange Orange Faint Pink Pink Helicobacter NoneVery Rare Rare Exist Many

-   3. Helicobacter pylori was suspended in physiological saline to give    about 10⁹ organisms per milliliter. 50 μL of this innoculum was    transferred to tubes containing 1 mL of citrate-phosphate buffer at    various concentrations such as ½, ¼, and 1/10 for 15 minutes, ½, 1/4    , and 1/10 for 30 minutes, and ½, ¼, and 1/10 for 45 minutes. Paired    tubes were innoculated with and without 6 mM Urea. Following a 30    minute room temperature incubation, the suspensions were subcultured    on agar plates using a 1 μL loop. These plates were substantially    free of bismuth and bile acids. Plates were incubated    microaerophilically at 37° C. for 72 hours. This procedure is    illustrated in FIG. 11.

Table 9 shows urease test results following 72 hours of growth of H.pylori on media prepared with dilutions of UDCA (000112B-1) bismuthcitrate (OSABY) or both UTDCA and bismuth citrate. As indicated, longerexposure times increased the adverse effect of the solutions on H.pylori.

TABLE 9 Incubation Urease Test (min.) Dilution Time (min.) 1 30 60 120240 000112B-1 Control P P P P P 1/2 15 Y Y Y Y Y 30 Y Y Y Y Y 45 Y Y Y YY 1/4 15 Y FO FP P P 30 Y Y FO FO FO 45 Y Y Y Y Y 1/10 15 Y FO FO FO FO30 Y FO FO O P 45 Y Y Y FO FO OSABY Control P P P P P 1/2 15 Y Y Y Y Y30 Y Y Y Y Y 45 Y Y Y Y Y 1/4 15 Y Y Y Y Y 30 Y Y Y Y Y 45 Y Y Y Y Y1/10 15 Y FO FO FO P 30 Y Y Y Y Y 45 Y Y Y Y Y 000122B-1 + OSABY ControlP P P P P ½ 15 Y Y Y Y Y 30 Y Y Y Y Y 45 Y Y Y Y Y 1/4 15 Y Y Y Y Y 30 YY Y Y Y 45 Y Y Y Y Y 1/10 15 Y Y Y FO FO 30 Y FO FO FO P 45 Y Y Y Y Y

1. An aqueous solution free of precipitates or particles comprising: (i)a first material selected from the group consisting of an aqueoussoluble bile acid salt, a bile acid conjugated with an amine by an amidelinkage, and combinations thereof; (ii) a second material selected fromthe group consisting of maltodextrin, dextrin, corn syrup, corn syrupsolid, soluble starch, and dextrans, and combinations thereof; (iii) atleast one alkali, and (iv) water, wherein the first and second materialsboth remain in solution for all pH values obtainable in an aqueoussystem.
 2. The solution of claim 1, wherein the solution is selectedfrom the group consisting of a syrup and a thick syrup.
 3. The solutionof claim 1, wherein the bile acid portion of said first material isselected from the group consisting of ursodeoxycholic acid,chenodeoxycholic acid, cholic acid, hyodeoxycholic acid, deoxycholicacid, 7-oxolithocholic acid, lithocholic acid, iododeoxycholic acid,iocholic acid, tauroursodeoxycholic acid, taurochenodeoxycholic acid,taurodeoxcycholic acid, glycoursodeoxycholie acid, tauroeholic acid, andglycocholic acid.
 4. The solution of claim 1, wherein the solutioncomprises one or more additional bile acid salts and amine-conjugatedbile acids conjugated by an amide linkage.
 5. The solution of claim 1further comprising an agent having anti-inflammatory activity.
 6. Thesolution of claim 1 further comprising an agent having analgesicactivity.
 7. The solution of claim 1 further comprising an agent havinganticonvulsant activity.
 8. The solution of claim 1 further comprisingan agent capable of prolonging survival time in hypoxic conditions. 9.The solution of claim 1 further comprising an agent capable ofalleviating or ameliorating a condition selected from the groupconsisting of stomatitis, gingivoglossitis and toothache.
 10. Thesolution of claim 1, wherein the aqueous solution is free of allprecipitates or particles.
 11. An aqueous solution free of precipitatesor particles comprising: (a) a first material selected from the groupconsisting of an ursodeoxycholic acid salt, an ursodeoxycholic acidconjugated with an amine by an amide linkage, and combinations thereof;(b) a second material which is maltodextrin; (c) at least one alkali,and (d) water, wherein the first and second materials both remain insolution for all pH values obtainable in an aqueous system and whereinthe weight ratio of the second material to the first material is about25:1.
 12. The solution of claim 11, wherein the aqueous solution is freeof all precipitates or particles.
 13. An aqueous solution free ofprecipitates or particles comprising: (i) a first material selected fromthe group consisting of an aqueous soluble bile acid salt, a bile acidconjugated with an amine by an amide linkage, and combinations thereof;(ii) a second material selected from the group consisting ofmaltodextrin, dextrin, corn syrup, corn syrup solid, soluble starch, anddextrans, and combinations thereof; and (iii) water, wherein the firstand second materials both remain in solution for all pH valuesobtainable in an aqueous system.
 14. The solution of claim 13, whereinthe aqueous solution is free of all precipitates or particles.